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微生物细胞在微通道表面的共价固定化。

Covalent Immobilization of Microbial Cells on Microchannel Surfaces.

机构信息

Department of Medical Biochemistry and Biophysics, Umeå University, Umeå, Sweden.

Faculty of Chemistry and Chemical Technology, University of Ljubljana, Ljubljana, Slovenia.

出版信息

Methods Mol Biol. 2020;2100:417-426. doi: 10.1007/978-1-0716-0215-7_28.

Abstract

Microfluidic devices with integrated biological material have found many applications in analytics (e.g., protein and DNA analysis), biochemistry (e.g., PCR), and medical diagnostics (e.g., ELISA test). Recently they are also considered as promising tools for bioprocess development and intensification. In order to enable long-term biocatalyst use and to facilitate its separation from the product, immobilization within the microreactor is often preferred over the use of free enzymes or cells. Surface immobilization is frequently selected due to the very high surface-to-volume ratio of microfluidic devices that offers the possibility for high biocatalyst load and at the same time good biocatalyst accessibility. Moreover, such reactor design prevents the increase in backpressure, often encountered in packed-bed or monolithic microreactors.Microbial cells are beneficial over the isolated enzymes in many biotransformations, especially in multistep syntheses and in cofactor-dependent reactions. Their immobilization within microreactors, especially made from disposable polymers, is of a big interest for analytical and synthetic applications.This chapter describes procedure for immobilization of eukaryotic and prokaryotic cells onto inner surfaces of microreactors made from various polymeric materials and glass. Cells could be immobilized in high densities and remain stably attached over several days of continuous microreactor operation.

摘要

微流控设备与集成的生物材料在分析学(如蛋白质和 DNA 分析)、生物化学(如 PCR)和医学诊断(如 ELISA 测试)中有许多应用。最近,它们也被认为是生物工艺开发和强化的有前途的工具。为了能够长期使用生物催化剂并促进其与产物的分离,通常优先在微反应器内进行固定化,而不是使用游离酶或细胞。由于微流控设备具有非常高的表面积与体积比,这为高生物催化剂负载提供了可能性,同时也保证了良好的生物催化剂可及性,因此表面固定化经常被选择。此外,这种反应器设计可以防止在填充床或整体式微反应器中经常遇到的背压增加。在许多生物转化中,微生物细胞比分离的酶更有益,特别是在多步合成和依赖辅因子的反应中。它们在微反应器中的固定化,特别是由一次性聚合物制成的微反应器,对于分析和合成应用具有很大的兴趣。这一章描述了将真核和原核细胞固定在由各种聚合物材料和玻璃制成的微反应器内部表面的程序。细胞可以以高密度固定化,并且在连续微反应器操作数天内仍稳定附着。

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